Dataset Information

Double lysis: an integrative time-saving method yielding high-quality RNA from strawberry.

ABSTRACT: BACKGROUND:The isolation of high-quality RNA from strawberry leaves and fruits is notoriously cumbersome. Both tissues are extremely rich in active secondary metabolites, as phenolics and pigments that inevitably perturb the isolation of RNA. Many protocols have been developed to address this problem. However, they are either costly, or time-consuming, in particular for high number of many plant samples. We describe here a new method with an easy-to-handle approach to obtain high-quality RNA from strawberry leaves and fruits. The method, referred to as double lysis, uses a novel combination of CTAB and Trizol protocols. RESULTS:Compared to conventional Trizol-dependent protocols, either used individually, or in a commercial spin-column kit, the new method yields RNA at lower costs, but of significantly improved quality. The RNA obtained by double lysis was very pure as indicated by 260/280 ratios of 2.06 (leaves) and 2.07 (fruits), while 260/230 ratio was 2.07 and 1.75, respectively. Also visually, RNA sediments from double lysis showed a white color, indicative of efficient elimination of polyphenolics and pigments. In contrast, traditional Trizol method produced reddish precipitates. The purity of RNA isolated by double lysis enabled successful removal of genomic DNA and, thus, allowed for more efficient cDNA synthesis and RT-PCR. In addition, the suggested method is able to yield RNA with fully preserved integrity from strawberry leaves, fruits in addition to petals and roots. CONCLUSION:Double lysis is a new RNA isolation protocol developed through the integrative coupling of CTAB and Trizol reagents. The method is cost-effective, robust, time-saving, and can cope even with recalcitrant plant tissues with high contents of phenolics and pigments, such as strawberry leaves and fruits.

SUBMITTER: Hazman M

PROVIDER: S-EPMC7311625 | biostudies-literature | 2020 Jun

REPOSITORIES: biostudies-literature

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Double lysis: an integrative time-saving method yielding high-quality RNA from strawberry.

Hazman Mohamed M Kabil Farida F Abd Elhamid Shrouk S Nick Peter P

Journal, genetic engineering & biotechnology 20200624 1

<h4>Background</h4>The isolation of high-quality RNA from strawberry leaves and fruits is notoriously cumbersome. Both tissues are extremely rich in active secondary metabolites, as phenolics and pigments that inevitably perturb the isolation of RNA. Many protocols have been developed to address this problem. However, they are either costly, or time-consuming, in particular for high number of many plant samples. We describe here a new method with an easy-to-handle approach to obtain high-quality ...[more]

PMID: 32577923

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Project description:Reduction of radiation dose in x-ray imaging has been recognized as a high priority in the medical community. Here the authors show that a regional adaptive exposure method can reduce dose-area product (DAP) in x-ray fluoroscopy. The authors' method is particularly geared toward providing dose savings for the pediatric population.The scanning beam digital x-ray system uses a large-area x-ray source with 8000 focal spots in combination with a small photon-counting detector. An imaging frame is obtained by acquiring and reconstructing up to 8000 detector images, each viewing only a small portion of the patient. Regional adaptive exposure was implemented by varying the exposure of the detector images depending on the local opacity of the object. A family of phantoms ranging in size from infant to obese adult was imaged in anteroposterior view with and without adaptive exposure. The DAP delivered to each phantom was measured in each case, and noise performance was compared by generating noise arrays to represent regional noise in the images. These noise arrays were generated by dividing the image into regions of about 6 mm(2), calculating the relative noise in each region, and placing the relative noise value of each region in a one-dimensional array (noise array) sorted from highest to lowest. Dose-area product savings were calculated as the difference between the ratio of DAP with adaptive exposure to DAP without adaptive exposure. The authors modified this value by a correction factor that matches the noise arrays where relative noise is the highest to report a final dose-area product savings.The average dose-area product saving across the phantom family was (42 ± 8)% with the highest dose-area product saving in the child-sized phantom (50%) and the lowest in the phantom mimicking an obese adult (23%).Phantom measurements indicate that a regional adaptive exposure method can produce large DAP savings without compromising the noise performance in the image regions with highest noise.

Dataset Information

Double lysis: an integrative time-saving method yielding high-quality RNA from strawberry.

Publications

Double lysis: an integrative time-saving method yielding high-quality RNA from strawberry.

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